Natural products may increase cytotoxic activity of Natural Killer Cells (NK) Tumor Necrosis Factor alpha (TNF-a) while decreasing DNA damage in patients with late-stage cancer. Pilot studies have suggested that a combination of Nutraceuticals can raise Natural Killer cell function and TNF-a alpha activity and result in improved clinical outcomes in patients with late stage cancer. The objective of the study is to determine if Nutracxeuticals can significantly raise Natural Killer cell function and Tumor Necrosis Factor levels in patients with late stage cancer. After informed consent was obtained, 20 patients with stage IV, end-stage cancer were evaluated (one bladder, five breast cancer, two prostate cancer, one neuroblastoma, two non-small cell lung, three colon cancer, 1 mesothelioma, two lymphoma, one ovarian cancer, one gastric, one osteosarcoma). Transfer Factor Plus (TFP, 3 tablets 3 times per day), IMUPlus (non denatured mild whey protein, 40 gm/day); Intravenous (50 to 100 gm/day) and oral (1-2 gm/day) ascorbic acid; Agaricus Blazeii Murill teas (10 gm/day); Immune Modulator Mix (a combination of vitamin, minerals, antioxidants and immune-enhancing natural products); nitrogenated soy extract (high levels of genistein and dadzein) and Andrographis Paniculata (500 mg twice daily) were used. Baseline NK function by standard 4 h ⁵¹Cr release assay and TNF alpha and receptor levels were measured by ELISA from resting and phytohemagglutinin (PHA) stimulated adherent and non-adherent Peripheral Blood Mononuclear Cell (PBMC). Total mercaptans and glutathione in plasma were taken and compared to levels measured 6 months later. Complese blood counts and chemistry panels were routinely monitored. As of a mean of 6 months, 16/20 patients were still alive. The 16 survivors had significantly higher NK function than baseline (p<.01 for each) and TNF-a levels in all four cell populations studied (p<.01 for each). Total merceptans (p<.01) and TNF-a receptor levels were significantly reduced (p<.01). It was also observed that hemoglobin, hematocrit and glutathione levels were significantly elevated. The only toxicity noted was occasional diarrhea and nausea. The quality of life improved for all survivors by SF-36 form evaluation. An aggressive combination of immunoactive Nutraceuticals was effective in significantly increasing NK function, other immune parameters and hemoglobin from PBMC or plasma in patients with late stage cancers. Nutraceutical combinations may be effective in late stage cancers. Clinical outcomes evaluations are ongoing.

Introduction

Despite enormous expenditures, little real progress has been made in the treatment of most cancers. Cancer incidence in the United States has risen 60 percent since 1950 and 27 fold since 1900 according to Surveillance, Epidemiology, and End Results (SEER). ^[1] Each year one in two men and one in three women in the U.S. develop some form of cancer according to the American Cancer Society.^[1] Mortality has climbed dramatically. In 1999, 560,000 people died of cancer in the U.S. alone. The mortality rate for cancer in 1994 was 6 percent higher than in 1970.^[1] Major gains have been made using chemotherapy in childhood leukemias, testicular cancer and some rare cancers, but the ability of chemotherapy and radiation to alter mortality of most cancers is negligible at best. Because of these facts, many Americans are utilizing some sort of alternative medicine in their battle against cancer. Estimates range from 9% of patients by the National Cancer Institute (NCI) to 60% of patients by other estimates. It appears that only about 5% of patients completely abandon traditional treatments for alternatives.^[2] There is a significant need for research to determine, which if any alternative therapies show promise in the treatment of cancers. The role of the immune system in fighting cancer was first proposed by Paul Ehrlich around 1900, and the concept renewed by Lewis Thomas in 1959 and MacFarlane Burnet in 1969. Various research centers have been looking at immunotherapy for some time, and in fact in certain cancers recombitant interleukin-2 (IL-2) and interferons are used, but these therapies are plagued with serious side-effects. Various natural products have been shown to have immune modulating properties.^[3]

The importance of the immune system in cancer prevention and adjuvant therapy has been well-established. Penn reports the following evidences for the role of the immune system in the development of cancers.^[4]

1. Children with immunodeficiency diseases have increased rates of lymphoma, leukemia and Hodgkin's disease.

2. High rates of Kaposi's sarcoma and lymphoma in HIV infected individuals.

3. In organ transplant patients on immunosuppressive medications, there is a 3 fold increase in malignancies including skin, lips, vulva, anus, liver, lymphoma and Kaposi's sarcoma.

4. Cancer risk increases with duration of immunosuppressive treatmen. In a study of heart transplant patients, cancer incidence increased 3 fold after one year of immunosuppressive therapy and 26 fold after 5 y.

5. Patients with autoimmune diseases treated with immunosuppressive therapy showed increased incidence of acute leukemia, lymphoma, liver cancer, bladder cancer and skin cancer.

6. Secondary tumores are common in cancer patients who receive immunosuppressive chemotherapy treatment. These may include acute leukemias, lymphoma, and bladder cancers. However this increase may be due to the DNA damage caused by the chemotherapeutic agent.

The importance of Natural Killer (NK) cells in effective immunotherapy has been broadly accepted. Our laboratory has consistently found that stage 4 cancer patients typically have severely depressed NK function, and sometimes decreased numbers (unpublished data). This is consistent with a plethora of other research on the subject.^[5] Decreases in NK function are believed to be, at least in part, attributable to neoplasm's ability to "steer" the immune system towards T-helper 2 (Th2) vs. cell mediated or T-helper 1 (Th1) immunity. Many cancers secrete interleukin-10 (IL-10) as part of this mechanism.

Human Tumor Necrosis Factor-alpha (TNF-a) is a 233 amino acid residue, nonglycosylated polypeptide that exists as either a transmembrane or soluble protein. When expressed as a 26 kDa membrane bound protein, TNF-a consists of a 29 amino acid residue cytoplasmic domain, a 28 amino acid residue transmembrane segment, and a 176 amino acid residue extracellular region. The soluble protein is a 17 kDa, 157 amino acid residue molecule that normally circulates as a homotrimer. The variety of cell types known to express TNF-alpha includes macrophages, CD4⁺ and CD8⁺ T cells adipocytes, keratinocytes, mammary and colon epithelium, osteoblasts mast cells, dendritic cells, pancreatic beta-cells, astrocytes, neurons, monocytes, and steroid-producing cells of the adrenal zona reticularis.^[6] Results of adenoviral mediated gene transfer of human TNF-alpha indicate a reduced systemic toxicity with an enhanced local antitumor effect in mice.^[7] Gillio-Tos and colleagues demonstrated that TNF-alpha antitumoral effect was due to apoptosis mediated by the down-regulation of the antiapoptotic gene bcl-2.^[8] Also, retroviral mediated gene transfer of the human TNF-alpha (hTNF) gene into U373MG human glioblastoma cells was shown to result in a reduction of the cells growth rate as compared to the parental cells.^[9]

Tumor necrosis factor has direct in vitro antitumor activity on 30 to 50% of cell lines. Because of its pleiotropy, it is not fully understood which mechanism is responsible for its necrotic and apoptotic activities. When membrane bound, TNF-alpha exhibits antineoplastic potential, but when free floating, is a negative prognosticator of survival, being associated with wasting both in cancer and AIDS patients.

Transfer factors are small peptides that act immune system regulatory cytokines.^[10] Transfer Factor Plus (TFP) contains several immunoactive components that have been shown to act synergistically in raising NK function and also effective as adjuvant therapy in cancer therapy.^[11] In fact, one study (subitted) has suggested that Transfer Factor Plus alone may raise Natural Killer function by nearly 250%. One component, inositol hexaphosphate (IP-6), has been shown to have an at least thee anti-cancer properties (NK enhancer, induces p53 and diminishes mutagenesis). The other components, namely, three medicinal mushrooms, mannans, thymic lipoproteins, and beta glucan, have all benn proven to enhance Natural Killer function as well as transfer factor itself.

Non-denatured milk whey protein isolates have been found to have a number of bioactive properties. Bovine milk contains around 3% protein by weight, with only 6.3% of that protein being whey protein. Normal processes to separate the whey component from the other constituents leads to significant denaturing of the bioactive whey proteins. Non-denatured whey protein isolates utilize proprietary processes to attain a protein containing over 90% non-denatured whey protein. Bioactive componenets include lactoferrin, lysozyme, lactoperoxidase, glycomacropeptide, alpha-lactalbumin, and bovine-serum albumin. Lactoferrin exhibits anticancer, antiviral, antibacterial, and antifungal activity. It plays an active role in iron transport, and active role in the cytotoxic defenses of neutrophils, and scavenges free iron which acts as a free radical.^{[12, 13]}

Of particular interest with non-denatured milk whey proteins, is their up-regulation of intracellular glutathione via supplying cystine. The roles of glutathione (GSH) has been summarized by Gutman^[14] and include enhancement of immune function, elimination of toxins, elimination of carcinogens, antioxidant cell protection, protection from ionizing radiation, DNA synthesis and repair, protein synthesis, prostaglandin synthesis, leukotriene syntheses, amino acid transport and enzyme activity and regulation. Bounous et al have demonstrated that non-denatured whey protein isolate formula increased lymphocyte intracellular glutathione by greater than 120% in mice vs. mice fed standard, commercially available whey protein concentrates, or casein proteins. ^{[15, 16]}

Table I. Ingredients in Immune Enhancing Formula

Kennedy demonstrated that non-denatured whey protein isolate formula increased glutathione in normal cells, but decreased glutathione in cancer cells.^[17] It has been proposed that cancer cells produce more intracellular glutathione than normal cells in order to protect themselves from various reactive oxygen species. Thus, it would appear possible that the ingestion of non-denatured whey protein isolates may oxidatively stress cancer cells, while protecting normal cells. Bounous et al. have also demonstrated that mice fed non-denature whey protein isolates exhibit a significantly smaller tumor load than controls, when both groups are fed know carcinogens.^[18]

Additionally, wasting (cachexia) can be a significant problem for the cancer patient. Though the mechanisms of cachexia are not fully understood, it is clear that there are major metabolic alterations in the cancer patient.^[19] Tumor cells usually resort to the anaerobic rather than the aerobic metabolism for the production of energy. This type of metabolism is grossly inefficient and generates large amounts of lactic acid that must in turn be regenerated back into glucose in the liver, via the cori cycle, which also requires additional amounts of energy. Eventually the body begins to consume skeletal muscle for energy. Cachexia is also characterized by decreased appetite. non-denatured whey protein isolate supplies a concentrated form of easily digestible high efficiency protein, without the need to consume large volumes of food. Finally, a number of clinics, including our own, have observed that patients rendered anemic due to chemotherapy and radiation appear to have robust recovery of their hemoglobin and hematocrit when fed adequate levels of non-denatured whey protein isolate (personal communication).

Our institute employs the use of a specialized powder containing a number of immune enhancing ingredients, including high potency antioxidants (Vitamins A, C, E, Selenium), products that enhance phase 2 detoxification in the liver, and products known to have effects on restoring cell cycle dynamics in cancer cells via a number of mechanisms including upregulation of tumor suppressor genes, downregulation of oncogenes, and other modulating effects on cyclin dependent kinases and other modulators of cell cycle dynamics (green tea extract, andrographis paniculate, Vitamin A, Vitamin D, to name a few).^[20] The full regime each patient used is shown in Table 2.

Table II. Regime

1. Nutraceuticals: 
   ・ Transfer Factor Plus, ImuPlus milk whey protein, Agaricus Blazeil Murill teas, Transfer Factor Plus, Beta Glucan, immune augmenting powder, and Soy Extract (and others cancer specific e.g., PC-Spes for Prostate)

2. Hyperthermia-local and systemic (far infrared sauna)
3. Light generator
4. Filtered, magnetic resonant water
5. Enemas
6. Lymphatic massage
7. Immunoimagery
8. Vegetarian, low sugar diet
9. Digestive enzymes
10. DMSA
11. Intravenous semi-benzyl Ted ascorbic acid (50-10 gm/day)

The purpose of this pilot study is to show if a regimen of natural immunomodulators could increase Natural Killer function and augment TNF-a production from PBMC; decrease tumor load; and increase quality of life in end-stage patients with cancer.

Alternative Cancer Treatment and Natural Killer cell and TNF-alpha Function

Results on the Laboratory Studies on NK cell and TNF-a function in relation to Cancer Treatment

Discussion on Alternative Cancer Treatments' Effectiveness on Natural Killer Cell and TNF-alpha Function

Natural Killer Cells in Human Cancer

Immune System & Diseases

Transfer Factor & Immune Function that affect Cancer

References

^[1] SEER (Surveillance, Epidemiology, and End Results) Cancer Statistic Review, National Cancer Institute, 1996
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^[2] Boik J. Cancer and Natural Medicine, Oregon Medical Press, Princeton, Minnesota, 1996, 3
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^[3] Moss R. Cancer Therapy: The Independent Consumer's Guide, Equinox Press, Brooklyn, 1996
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^[4] Korbelik M., Sun J., Cancer Treatment by Photodynamic Therapy Combined with Adoptive Immunotherapy using Genetically Altered Natural Killer Cell Line, Int. J. Cancer 93 (2001) 269-274
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^[5] Miki T., Yano S., Hanibuchi M., Sone S., Bone Metastases Model with Multiorgan Dissemination of Human Small Cell Lung Cancer (SBC-5) Cells in Natural Killer Cell Delpleted Mice, Oncol. Res., 12 (2001) 209-217
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^[6] Zhang M., Tracey K. Tumor Necrosis Factor, The Cytokine Handbook, 3, Academic press Ltd., 1998, pp. 517-548
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^[7] Marr R., Hitt M., Muller WJ., Gauldie J., Graham FL., Tumour Therapy in Mice using Adenovirus Vectors Expressing Human TNFa, J. Oncol., 12 (1998) 509-515
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^[8] Gillio-Tos A., Cignetti A., Rovera G., Foa R., Retroviral Vector-Mediated Transfer of the Tumor Necrosis Factor Alpha Gene into Human Cancer Cells Restores and Apoptotic Cell Death Program and Induces a Bystander-killing Effect, Blood, 87 (1996) 2486-2495.
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^[9] Walther W., Stein U., Pfeil D., Gene Transfer of Human TNF Alpha into Glioblastoma Cells Permits Modulation of mdr1 Expression and Potentiation of Chemosensitivity, Int. J. Cancer, 61 (1995) 832-839
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^[10] Kirkpatrick C.H., Activity and Characteristics of Transfer Factors, Biotherapy, 9 (1996) 13-16
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^[11] Bomford D., Moreno J., Mechanisms of the Anti-tumor Effect of Glucans and Fructosans: A Comparison with C, Parvum. Br. J. Cancer, 36 (1997) 4148-4153
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^[12] Kawasaki M., Inhibitin of Kappa-casein Glycomacropeptide and Lactoferrin of Influenza Virus Hemagglutination, Int. J. Cancer, 57 (1993) 1214-1215
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^[13] Marchetti J., Metal Complexex of Bovine Lactoferrin Inhibit in Vitro Replication of Herpes Simplex Virus Type 1 and 2, BioMetals, 11 (1994) 89-94
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^[14] Gutman J., Schettini S. The Ultimate GSH Handbook, Gutman and Schettini, Montreal, 1998
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^[15] Baruchel S., Viav G., Olivier R., Bounous G., Wainberg M. Nutraceutical Modulation of GSH with a Humanized Native Milk Serum Protein Isolate, Immunocal: Application AIDS and Cancer is Oxidative Stress and Redox Regulation: Cellular Signaling, AIDS, Cancer and other Diseases. Symposium, Institute Pastuer, 1996,
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^[16] Bounous G., Gold P., The Biological Activity of Undenatured Whey Proteins: Role of Glutathione, Clin. Invest Med., 14 (1991) 296-309
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^[17] Kennedy R.S., Konok G.P., Bounous G., Baruchel S., Lee TD., The Use of a Whey Protein Concentrate in the Treatment of Patients with Metastatic Carcinoma: A Phase I-II Clinical Study, Anticancer Res., 15 (1995) 2643-2649
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^[18] Bounous G., Papenburg R., Kongshavn P.A., Gold P., Fleiszer D., Dietary Whey Protein Inhibits the Development of Dimethylhydrazine Induced Malignancy, Clin. Invest Med., 11 (1998) 213-217
           [1]

^[20] Georgiev V.N., Durnev A.D., Seredenin S.B., [?] Ubiquinones and the Body's Antimutagenic Defense, Voprosy Meditsinskoi Khimii, 40 (1994) 8-9
           [1]

Transfer Factor can strengthen your immune cells (NK cells) by educating them to recognise harmful invasion to your body, remember the past invasion and respond accordingly with the best possible way.  NK cells are known for the important roll in the fight against cancer.